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G aphical abs ac
pp xxx–xxxSyn hesis o subs i u ed 2-(b-
DD
-glucopy anosyl)-benzimidazoles and hei e alua ion as inhibi o s o glycogen
phospho ylase
É a Boko , Enik}
o Szilágyi, Tibo Docsa, Pa
´l Ge gely, La
´szlo
´Somsa
´k
*
CAR 6384 No. o Pages 1, Model 5G
5 Feb ua y 2013
Highligh s
"Syn hesis o C-(b-
DD
-glucopy anosyl) o mimida es and - hio o mimida es. "New syn hesis o C-(b-
DD
-glucopy anosyl)-benzimidazoles.
"Low mic omola inhibi o s o glycogen phospho ylase.
X=O,S
O
OR
1
R
1
O
R
1
O
OR
1
N
H
N
O
OBz
BzO
BzO COOH
OBz
O
OBz
BzO
BzO CXNH
2
OBz
O
OBz
BzO
BzO C
OBz NH
XE
H
2
N
H
2
N
R
2
E
3
OBF
4
om
X=O
R
2
R
1
=Bz H
NO
2
O
OH
HO
HO
OH
N
H
N
bes inhibi o
K
i
=2.1μM( abbi mu
s
R
2
=5-F,5-Cl,5-B ,5-
N
4-Me, 5-Me, 5,6-di
1
Syn hesis o subs i u ed 2-(b-D-glucopy anosyl)-benzimidazoles and
hei e alua ion as inhibi o s o glycogen phospho ylase
É a Boko
a
,Enik}
oSzilágyi
a
,Tibo Docsa
b
,Pál Ge gely
b
,László Somsák
a,
⇑
a
Depa men o O ganic Chemis y, Uni e si y o Deb ecen, POB 20, H-4010 Deb ecen, Hunga y
b
Depa men o Medical Chemis y, Medical and Heal h Science Cen e, Uni e si y o Deb ecen, Egye em é 1, H-4032 Deb ecen, Hunga y
a icle in o
A icle his o y:
Recei ed 3 No embe 2012
Recei ed in e ised o m 15 Janua y 2013
Accep ed 16 Janua y 2013
A ailable online xxxx
Keywo ds:
C-Glycopy anosyl- o mimida e
C-Glycopy anosyl- hio o mimida e
2-C-Glycopy anosyl benzimidazole
Glycogen phospho ylase
Inhibi o
abs ac
Mic owa e assis ed condensa ion o O-pe benzoyla ed C-(b-D-glucopy anosyl) o mic acid wi h
1,2-diaminobenzenes in he p esence o iphenylphosphi e ga e he co esponding O-p o ec ed
2-(b-D-glucopy anosyl)-benzimidazoles in mode a e yields. O-Pe benzoyla ed C-(b-D-glucopy anosyl)
o mamide and - hio o mamide we e ans o med in o he co esponding e hyl C-(b-D-glucopy anosyl)
o mimida e and - hio o mimida e, espec i ely, by E
3
OBF
4
. T ea men o he o mimida e wi h
1,2-diaminobenzenes a o ded O-p o ec ed 2-(b-D-glucopy anosyl)-benzimidazoles-in good o excellen
yields. Simila eac ion o he hio o mimida e ga e hese compounds in lowe yields. The O-benzoyl
p o ec ing g oups we e emo ed by he Zemplén p o ocol. These es compounds we e assayed agains
abbi muscle glycogen phospho ylase (GP) b, he p o o ype o li e GP, he a e limi ing enzyme o
glycogen deg ada ion. The bes inhibi o s we e 2-(b-D-glucopy anosyl)-4-me hyl-benzimidazole
(K
i
= 2.8
l
M) and 2-(b-D-glucopy anosyl)-naph ho[2,3-d]imidazole (K
i
= 2.1
l
M) exhibi ing a 3–4 imes
s onge binding han he unsubs i u ed pa en compound.
Ó2013 Published by Else ie L d.
1. In oduc ion
Inhibi ion o glycogen phospho ylase (GP) has been conside ed
as an e ec i e he apeu ic app oach in comba ing ype 2 diabe es
( o he biochemical and pha macological backg ound o a ge ing
li e GP as a alida ed concep o lowe ing blood glucose le els,
please su ey ecen e iew a icles
1–6
). Fu he mo e, he pha ma-
ceu ical u ili y o GP inhibi o s in he in e en ion o o he
diseased s a es associa ed wi h GP ac i i y (e.g., ca dio ascula dis-
50
o de s,
7–9
ischaemic lesions,
10–13
and umo ous g ow h
11,14–16
) has
also been unde in es iga ion.
A la ge a ay o compounds we e shown o ha e an inhibi o y
e ec agains his enzyme
17–19
including glucose de i a i es
20,21
which p ima ily bind o he ca aly ic si e o he enzyme. In he
cou se o sea ching o po en glucose based inhibi o s, se e al
C-b-
D
-glucopy anosyl he e ocycles (Cha 1), such as e azole
22
1A, 1,3,4-oxadiazoles
22–24
1B, 1,2,4-oxadiazoles
23,25,26
1C,D, benzo-
hiazole
22
1E and benzimidazole
22
1F ha e been syn hesized and
some o hem p o ed o be e ficien agains abbi muscle glycogen
60
phospho ylase b(RMGPb, he p o o ype o GP enzymes o enzy-
ma ic es s
2
). In his class, 2-(b-
D
-glucopy anosyl)-benzimidazole
(1F) was he fi s compound o ha e a K
i
alue in he low mic omo-
la ange.
22,27
As e idenced by X- ay c ys allog aphy, he s ong
binding in he ca aly ic cen e is he esul o di ec and wa e
media ed H-bonds be ween he p o ein and he he e oa oma ic
ing, and an de Waals in e ac ions o he la ge a oma ic pa in
he so-called b-channel o he enzyme.
27
The highe a fini y o 1F
in compa ison wi h i s hio coun e pa 1E can be a ibu ed o
he di ec H-bond o he imidazole NH wi h he His377 main chain
70
ca bonyl g oup o he enzyme which is ce ainly absen o benzo-
hiazole 1E.
27
Analogous NH-(His377)CO in e ac ions we e iden i-
fied in o he GP enzyme-inhibi o complexes (e.g., in cases o
spi o( hio)hydan oin
28,29
and N-acyl-b-
D
-glucopy anosylamine
ype inhibi o s
30
) indica ing he p ominen impo ance o his spe-
cial H-b idge, as well. Addi ionally, X- ay c ys allog aphic in es i-
ga ion o 1F in complex wi h RMGPb e ealed ha besides he
ac i e si e he compound also occupied he new allos e ic si e,
and a new binding cle called he benzimidazole si e was also
disco e ed.
27
80
To ge an insigh in o he s uc u e–ac i i y ela ionship o his
ype o inhibi o he aim o ou p esen wo k has been o syn he-
size a se ies o subs i u ed 2-(b-
D
-glucopy anosyl)-benzimidazoles
and o e alua e hei e ec on RMGPb.
Fo he o ma ion o 2-C-glycosyl-benzimidazoles se e al syn-
he ic me hods a e known om he li e a u e mos o which ha e
been desc ibed o u anose based de i a i es. Fu anosyl benzimi-
dazoles we e p epa ed (a) by acid ca alysed condensa ion o C-gly-
co u anosyl o mic acids and o-phenylenediamine (OPD),
31,32
(b) by
coupling o C-glyco u anosyl o mic acids o hei chlo ides wi h
90
1,2-diaminobenzenes, ollowed by acid o POCl
3
media ed ing
closu e o he esul ing amide ype in e media es,
33,34
(c) in he
0008-6215/$ - see on ma e Ó2013 Published by Else ie L d.
h p://dx.doi.o g/10.1016/j.ca es.2013.01.011
⇑
Co esponding au ho . Tel.: +36 52512900x22348; ax: +36 52512744.
E-mail add ess: [email p o ec ed] (L. Somsák).
Q1
Q2
Q3
Ca bohyd a e Resea ch xxx (2013) xxx–xxx
Con en s lis s a ailable a SciVe se ScienceDi ec
Ca bohyd a e Resea ch
jou nal homepage: www.else ie .com/loca e/ca es
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011
eac ion o C-glyco u anosyl o maldoximoyl chlo ides (ac ually
he ni ile oxide ob ained by base induced dehyd ochlo ina ion)
wi h OPD,
35
(d) by in amolecula ing closu e o sui ably
p o ec ed 2-(pen i ol-1
0
-yl)-benzimidazoles ob ained om 2-li hi-
a ed benzimidazoles and suga lac one o acyclic pen ose de i a-
i es,
36–38
and (e) by acid induced cyclodehyd a ion o
unp o ec ed 2-( e i ol-1
0
-yl)-benzimidazoles.
39
Fo he syn hesis o 2-C-glycopy anosyl-benzimidazoles h ee
100
p ocedu es we e epo ed: (a) acid ca alysed condensa ion o
unp o ec ed C-glycopy anosylme hanals (gene a ed in si u om
he app op ia e dime hyl ace als) wi h OPD, ollowed by spon ane-
ous oxida ion o he in e media e benzimidazolines,
40
(b) eac ion
o O-pe benzoyla ed e hyl (C-b-
D
-glucopy anosyl) hio o mimida e
hyd ochlo ide (p epa ed om he co esponding glycosyl cyanide)
wi h OPD,
22
and (c) ea men o O-pe ace yla ed C-glycopy anosyl
o maldoximoyl chlo ides wi h OPD.
41
In addi ion, a 2-benzimidaz-
olyl moie y was also a ached o C-1 o
D
-galac al in he eac ion o
me hyl C-(2-deoxy-
D
-lyxo-hex-1-enopy anosyl) o mimida e and
110
he dihyd ochlo ide sal o OPD.
42
In his pape , we disclose u he syn he ic possibili ies o con-
s uc benzimidazole a he anome ic cen e o he py anose uni .
2. Resul s and discussion
2.1. Syn heses
C-(b-
D
-Glucopy anosyl) o mic acid 5, p epa ed om cyanide 2
ia amide 3
43
by he li e a u e p o ocol
44
(Table 1), was eac ed wi h
1,2-diaminobenzenes (a, ,h) in he p esence o iphenylphosphi e
in py idine unde mic owa e i adia ion (condi ions we e adap ed
om a published p ocedu e applied o non-suga based com-
120
pounds
45
). Al hough he con e sions we e comple e a 140 °Cin
20 min, he desi ed p oduc s could be isola ed only in mode a e
yields (8a: 45%, 8 : 53%, 8h: 43%). Ne e heless, i has o be no ed,
ha ea lie a emp s o ans o m 5in o benzimidazole 8a by he
classical acid ca alysed p ocedu e wi h con en ional hea ing
b ough abou no eac ion e en a ele a ed empe a u es.
22
The e o e, applica ion o he mo e eac i e iminoes e s 6and 7
was en isaged o he cons uc ion o benzimidazoles.
P e iously, hyd ochlo ide sal o O-pe benzoyla ed e hyl
C-(b-
D
-glucopy anosyl) hio o mimida e ob ained om glucopy -
130
anosyl cyanide 2by an acid ca alysed addi ion o E SH o he ni ile
g oup was used o his pu pose.
22
Since hese condi ions we e
a he unpleasan and subsequen ans o ma ion o he sal ga e
benzimidazole 8a in a 34% yield only, we se ou o p oduce and
use he ee o m o his ype o p ecu so . F ee hioimida e 7
was ob ained om hioamide 4by E
3
OBF
4
. The p epa a ion o
4
46
was also modified o a oid he use o H
2
S; hus, cyanide 2
was eac ed wi h P
4
S
10
in efluxing E OH as desc ibed o he syn-
hesis o alipha ic and a oma ic hioamides.
47
Simila ly o 7, imi-
da e 6was ob ained om amide 3by E
3
OBF
4
.
140
Nex , he p epa a ion o benzimidazole 8a om ei he 6o 7
was compa ed in p elimina y expe imen s. Reac ion o 6o 7wi h
OPD ga e 8a in 89% and 62% yields, espec i ely. The e o e, also
aking in o accoun he ins abili y o 7(decomposi ion was
obse ed on s o age a a e a ew days), u he eac ions o
ge he desi ed benzimidazoles we e pe o med wi h 6.
Simila ly o 8a, high yields we e also achie ed in eac ions o 6
wi h me hyl-subs i u ed 1,2-diaminobenzenes –h and 2,3-diami-
nonaph halene i, espec i ely (Table 1). A compa ison o he yields
o 8a, ,h ob ained om acid 5unde condi ions and om imi-
150
da e 6unde condi ions i, espec i ely, showed he ing closu e
o 6wi h 1,2-diaminobenzenes o be supe io o ha o 5.
On ea men o imida e 6wi h 1,2-diaminobenzenes con ain-
ing elec on wi hd awing subs i uen s (b–e) benzoic acid elimina-
ion was also obse ed, and beside he 2-glucopy anosyl
benzimidazoles 8b–e 4-subs i u ed-2-(3
0
,4
0
,6
0
- i-O-benzoyl-2
0
-
deoxy-
D
-a abino-hex-1-enopy anosyl)-benzimidazoles (1-C-benz-
imidazolyl glucals, 9b–e) we e also isola ed. Fu he mo e, o al
consump ion o he s a ing ma e ial 6in eac ion wi h
4-ni o-1,2-diaminobenzene (e) equi ed highe empe a u e, hus
160
cycliza ion was pe o med in boiling e hanol o yield benzimid-
azole 8e oge he wi h 9e. This endency o elimina ion migh
be a ibu ed o an inc eased acidi y o he C-1–H p o on in he
2-glucosyl-benzimidazoles p obably due o he p esence o he
elec on wi hd awing subs i uen in he a oma ic ing sys em.
Fo enzyma ic s udies emo al o he benzoyl p o ec ing g oups
o 8b–i was e ec ed by he Zemplén me hod o gi e es com-
pounds 10b–i in good yields.
The s uc u e o he new compounds was de e mined by
1
H and
13
C NMR spec oscopy. The p esence o a b-
D
-configu ed glucopy -
170
anosyl moie y in he
4
C
1
con o ma ion was confi med by he ici-
nal p o on–p o on coupling cons an s o compounds 6,7,8b–i
and 10b–i, espec i ely. In he
1
H NMR spec a o 1-C-he a yl glu-
cals 9b–e he obse ed small couplings be ween he icinal ing
p o ons (3–6 Hz) sugges ed a con o ma ional change o he py a-
nose ing ( om chai o hal chai ).
48
In he
13
C NMR spec a o
9b–e cha ac e is ic esonances appea ed o C-1
0
(143–145 ppm)
and C-2
0
(99–101 ppm) p o iding e idence o he unsa u a ed
na u e o he py anoid ing.
48
Among
13
C NMR signals o he benz-
imidazole uni s only hose o C-2 appea ed as sha p peaks a 146–
180
158 ppm, he o he s usually ga e b oad signals. These obse a ions
a e in acco d wi h he li e a u e expe iences,
35,41
and may be ex-
plained by he apid p o on exchange be ween N-1 and N-3 o
he benzimidazole moie y.
35,41,49
Addi ionally, in he
13
CNMR
spec a duplica ion o ce ain ca bon peaks was also obse ed (p i-
ma ily o he benzimidazole ca bon signals o example, in 8b,9c,
9e and 10b), which may e e o he coexis ence o he au ome ic
pai s. Fo he fluo o-benzimidazole 8b his phenomenon seemed
qui e clea as 14 esonances we e obse ed in he
13
C NMR spec-
um which could be en a i ely assigned on he basis o he cha -
190
ac e is ic C–F couplings
50
as illus a ed in Figu e 1.
O
OH
HO
HO C
OH
1
He R
He R K
i
[μM]
A
NN
NHN
- No inhibi ion
22
B
O
NN
R
CH
3
145
27
212
22
2-naph hyl 10 % a 625 μM
23
C
N
ON
R
2-naph hyl 38
25
D
N
NO
R
2-naph hyl 2.4
23
E
S
N
-76
27
229
22
F
N
H
N
-8.6
27
11
22
Cha 1. Inhibi o y po ency (K
i
)o C-glucosyl he e ocyclic de i a i es agains abbi
muscle glycogen phospho ylase b(RMGPb).
2É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011
2.2. Enzyme kine ic s udies
Enzyme kine ic measu emen s wi h abbi muscle glycogen
phospho ylase bwe e pe o med as desc ibed p e iously,
51,52
and he ob ained inhibi o cons an s a e gi en in Table 2. The
new compounds showed inhibi o y e ec s in he mic omola
ange and some o hem p o ed equi alen o somewha be e
inhibi o s han 10a (=1F in Cha 1).
Among benzimidazoles wi h a halogen subs i uen in he 5-po-
si ion fluo o de i a i e 10b exhibi ed modes ac i i y, while he
200
chlo o (10c) as well as b omo (10d) compounds we e as good
inhibi o s as 10a. Subs i u ion o he benzimidazole ing in he
same posi ion by a ni o g oup (10e) b ough abou a significan
dec ease o he inhibi ion. Among he alkyl subs i u ed de i a i es
he 5-me hyl- (10 ) and 4-me hyl-benzimidazoles (10g) displayed
simila and sligh ly be e (3- old inc ease) po ency han 10a,
espec i ely. Howe e , in oduc ion o wo me hyl g oups in o
he 5- and 6-posi ions o he he e ocycle (10h) esul ed in a
ema kable weakening o he inhibi ion. Finally, he naph ho[2,3-
d]imidazole 10i showed 4 imes s onge binding han 10a and
210
p o ed o be he mos e ec i e inhibi o o his se ies. To elucida e
he na u e o he binding modes o he new compounds o GP
enzyme X- ay c ys allog aphic s udies a e in p og ess and will be
epo ed elsewhe e.
Table 1
Syn hesis o 2-(b-D-glucopy anosyl)-benzimidazoles
3X=O(94%)
4X=S(66%)
+
6X=O(96%)
7X=S(90%)
8a-i R
1
=Bz 9a-i
a-i
5(76 %)
iii
i
i O
OR
1
R
1
O
R
1
O
OR
1
N
H
N
R
2
R
3
R
4
O
OBz
BzO
BzO COOH
OBz
O
OBz
BzO
BzO CXNH
2
OBz
O
OBz
BzO
BzO C
OBz NH
XE
O
BzO
BzO
OBz
N
H
N
R
2
R
3
R
4
10a-i R
1
=H
ii
2
O
OBz
BzO
BzO CN
OBz
H
2
N
H
2
N
R
2
R
3
R
4
i o 3
ii o 4
i) HB -AcOH, ;43 ii) P4S10, abs. E OH, e lux; iii) NO2, abs. CH2Cl2, ;44 i ) E 3O·BF4, abs. CH2Cl2,
A , ; ) P(OPh)3 (1.2 equi .), a o o h (1.0 equi ), abs. py idine, μW, 140 ºC, 20 min; i) a-i (2
equi .), abs. CH2Cl2, e lux; ii) ~1 M NaOMe in abs. MeOH, .
Reagen R
2
R
3
R
4
Condi ions and yields (%)
8910
aHH H 45 — ii —
c,d
i 89 ( om 6)—
i 62
a
( om 7)
bHF H i 46 35 ii 54
cHCl H i 76 24 ii 89
dHB H i 54 29 ii 58
eHNO
2
H i
b
46 20 ii 53
HMeH 53 — ii 87
i 83 —
gMe H H i 79 — ii 73
hHMeMe 43 — ii 78
i 81 —
iH i 79 — ii 77
a
Repo ed yield o 8a by he ea lie me hod om pe benzoyla ed b-
D
-glucopy anosyl cyanide ia he hioimida e hyd ochlo ide sal : 34%.
22
b
Modified eac ion condi ions we e necessa y o comple e consump ion o he s a ing ma e ial 6: abs. E OH, eflux.
c
Debenzoyla ion o 8a by he Zemplén p o ocol was ca ied ou ea lie . Repo ed yield o 10a: 84%.
22
d
Repo ed yield o 10a by a di e en me hod: 54%.
40
N
N
H
F
Bz
4
-β-D-Glc
p
(H)
1(3)
3(1) 3a(7a)
4(7)
5(6)
6(5)
7(4)
7a(3a)
2(2)
151.1 (s)
150.3 (s)
158.9 (d,
1
J
C-F
=237Hz)
158.2 (d,
1
J
C-F
=237Hz)
104.5 (d,
2
J
C-F
=24Hz)
98.0 (d,
2
J
C-F
=27Hz)
142.8 (d,
3
J
C-F
~9Hz)
134.2 (d,
3
J
C-F
~9Hz)
139.0 (s)
130.8 (s)
110.9 (d,
2
J
C-F
=23Hz)
109.7 (d,
2
J
C-F
=23Hz)
120.0 (d,
3
J
C-F
~7Hz)
112.3 (d,
3
J
C-F
~6Hz)
in e changable
assignmen s
Figu e 1. Ten a i e assignmen o
13
C NMR signals o au ome s o compound 8b.
É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx 3
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011

3. Conclusion
New syn he ic pa hways we e elabo a ed o he o ma ion o
subs i u ed 2-(b-
D
-glucopy anosyl)-benzimidazoles by he e ocyc-
liza ions o 2,6-anhyd o-aldonic acid de i a i es as p ecu so s.
Condensa ion o O-pe benzoyla ed C-(b-
D
-glucopy anosyl) o mic
acid wi h 1,2-diaminobenzenes in he p esence o P(OPh)
3
unde
220
mic owa e i adia ion ga e he desi ed benzimidazoles in mode -
a e yields. Ring closu e o O-pe benzoyla ed e hyl C-(b-
D
-glucopy -
anosyl) o mimida e, p epa ed om he co esponding o mamide
by E
3
OBF
4
, wi h he 1,2-diaminobenzenes a o ded a mo e
e ficien p ocedu e. Wi h elec on deple ed a oma ic diamines
undesi able b-elimina ion o benzoic acid om he 1,2-posi ions
o he glucopy anosyl moie y also ook place o yield 1-C-benzim-
idazolyl glucals besides he a ge compounds. Enzyme kine ic
s udies wi h he new 2-(b-
D
-glucopy anosyl)-benzimidazoles
showed he compounds o inhibi RMGPbin he low mic omola
230
ange. Subs i u ion o he benzimidazole moie y by a me hyl g oup
in he 4-posi ion o u he annela ion o a benzene ing p o ided
mo e e ficien inhibi o s han he unsubs i u ed pa en compound.
4. Expe imen al
4.1. Gene al me hods
Mel ing poin s we e measu ed on a Kofle ho -s age and a e
unco ec ed. Op ical o a ions we e de e mined wi h a Pe kin–El-
me 241 pola ime e a . NMR spec a we e eco ded wi h B uke
360 (360/90 MHz o
1
H/
13
C) spec ome e . Chemical shi s a e e -
e enced o Me
4
Si (
1
H), o o he esidual sol en signals (
13
C).
240
Mic owa e assis ed p ocedu es we e pe o med in a CEM-Disco e
Focused Mic owa e Syn hesis Sys em (2450 MHz) wi h a buil -in
in a ed empe a u e senso and a CEM-Explo e compu e con-
olled obo ic sample .
TLC was pe o med on DC-Alu olle Kieselgel 60 F
254
(Me ck),
and he pla es we e isualized unde UV ligh and by gen le
hea ing. Fo column ch oma og aphy Kieselgel 60 (Me ck,
pa icle size 0.063–0.200 mm) was used. Dichlo ome hane was
dis iled om P
4
O
10
and s o ed o e 4 Å molecula sie es.
2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl cyanide,
43
C-(2,3,4,
250
6- e a-O-benzoyl-b-
D
-glucopy anosyl) o mamide
43
and C-(2,3,4,
6- e a-O-benzoyl-b-
D
-glucopy anosyl) o mic acid
44
we e syn he-
sized acco ding o published p ocedu es.
4.2. C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl)
hio o mamide (4)
A solu ion o phospho us pen asulfide (2.23 g, 10 mmol) in
anhyd ous E OH (20 mL) was s i ed a o 1.5 h, hen 2,3,4,6- e -
a-O-benzoyl-b-
D
-glucopy anosyl cyanide
43
(2, 3.02 g, 5 mmol)
was added and he mix u e was hea ed a eflux empe a u e o
2h.A e cooling he he e ogenous mix u e o , he p ecipi a e
260
was fil e ed o and washed wi h E OH o yield 2.1 g (66%) o 4
as a pale yellowish solid. Mp: 199–201 °C (Li .
46
mp: 198–
200 °C).
1
H and
13
C NMR da a co espond o he epo ed spec a.
46
4.3. E hyl C-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)
o mimida e (6)
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) o mamide
43
(3,
1.0 g, 1.60 mmol) and E
3
OBF
4
(0.91 g, 4.80 mmol) we e dissol ed
in anhyd ous CH
2
Cl
2
(15 mL), he mix u e was s i ed a unde
A and moni o ed by TLC (1:1 E OAc–hexane). A e comple ion
o he eac ion (2 days), he mix u e was dilu ed wi h CH
2
Cl
2
270
(30 mL), ex ac ed wi h sa d aq NaHCO
3
solu ion (30 mL) and hen
wi h wa e (30 mL). The o ganic phase was d ied o e MgSO
4
,fil-
e ed and he sol en was e apo a ed. The c ude pale yellow amo -
phous p oduc (1.0 g, 96%) was used wi hou u he pu ifica ion.
R
: 0.55 (1:1 E OAc–hexane);
1
H NMR (CDCl
3
)d(ppm): 8.06–7.25
(21H, m, a oma ics, NH), 5.99, 5.72, 5.55 (3 1H, 3 pseudo ,
J= 10.6, 9.2 Hz in each, H-2, H-3, H-4), 4.70 (1H, dd, J= 11.9,
2.6 Hz, H-6a), 4.52 (1H, dd, J= 11.9, 5.3 Hz, H-6b), 4.25 (1H, ddd,
J= 9.2, 5.3, 2.6 Hz, H-5), 4.21 (1H, d, J= 9.2 Hz, H-1), 4.12–3.91
(2H, m, CH
2
), 0.84 (3H, , 6.6 Hz, CH
3
);
13
C NMR (CDCl
3
)d(ppm):
280
167.5, 166.1, 165.7, 165.1 (2) (CO, CNH), 133.5–128.2 (a oma ics),
75.8, 74.6, 73.6, 70.7, 69.1 (C-1–C-5), 62.8, 62.1 (C-6, CH
2
), 13.4
(CH
3
).
4.4. E hyl C-(2,3,4,6- e a-O-benzoyl-b-
D
-glucopy anosyl)
hio o mimida e (7)
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) hio o mamide
(4, 0.2 g, 0.31 mmol) and E
3
OBF
4
(0.18 g, 0.94 mmol) we e dis-
sol ed in anhyd ous CH
2
Cl
2
(4 mL), he mix u e was s i ed a
unde A and moni o ed by TLC (1:1 E OAc–hexane). A e comple-
ion o he eac ion (2 days), he mix u e was dilu ed wi h CH
2
Cl
2
290
(10 mL), ex ac ed wi h sa d aq NaHCO
3
solu ion (15 mL), hen
wi h wa e (15 mL). The o ganic phase was d ied o e MgSO
4
,fil-
e ed and he sol en was e apo a ed. The c ude pale yellow amo -
phous p oduc (0.19 g, 90%) was used wi hou u he pu ifica ion.
R
: 0.62 (1:1 E OAc–hexane);
1
H NMR (CDCl
3
)d(ppm): 8.06–7.24
(21H, m, a oma ics, NH), 5.95, 5.74, 5.70 (3 1H, 3 pseudo ,
J= 9.8, 9.1 Hz in each, H-2, H-3, H-4), 4.68 (1H, dd, J= 11.9,
2.8 Hz, H-6a), 4.51 (1H, dd, J= 11.9, 4.9 Hz, H-6b), 4.39 (1H, d,
J= 9.1 Hz, H-1), 4.22 (1H, ddd, J= 9.1, 4.9, 2.8 Hz, H-5), 2.88–2.71
(2H, m, CH
2
), 1.17 (3H, pseudo , 7.7, 7.0 Hz, CH
3
);
13
C NMR (CDCl
3
)
d(ppm): 172.7 (CNH), 166.1, 165.7, 165.1, 164.9 (CO), 133.4–128.2
(a oma ics), 80.6, 76.1, 73.9, 70.9, 69.1 (C-1–C-5), 62.8 (C-6), 23.1
(CH
2
), 13.1 (CH
3
).
4.5. Gene al p ocedu e I o he syn hesis o subs i u ed
2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-glucopy anosyl)-
benzimidazoles om O-pe benzoyla ed
C-(b-
D
-glucopy anosyl) o mic acid (5)
C-(2,3,4,6-Te a-O-benzoyl-b-
D
-glucopy anosyl) o mic acid
44
(5, 0.1 g, 0.16 mmol), a 1,2-diaminobenzene (ao o h, 0.16 mmol,
1equi ) and iphenyl phosphi e (50
l
l, 0.19 mmol, 1.2 equi )
310
we e dissol ed in anhyd ous py idine (2 mL). The closed ial was
i adia ed by mic owa es o 20 min a 140 °C (maximum p es-
Table 2
Kine ic da a on he inhibi ion o abbi muscle glycogen phospho ylase bby he new
compounds 10b–i
O
OH
HO
HO
OH
N
H
N
R
2
R
3
R
4
10 R
2
R
3
R
4
K
i
(
l
M)
aH H H 8.6
27
11
22
bHF H55
cH Cl H 9.7
dH B H 7.5
eHNO
2
H 179
HMeH12
gMe H H 2.8
hH Me Me 152
iH2.1
Q4
4É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011
su e: 20 ba , powe 220 W). The comple ion o he eac ion was
moni o ed by TLC (1:1 E OAc–hexane). The eac ion mix u e was
concen a ed unde educed p essu e, aces o py idine we e
emo ed by epea ed co-e apo a ions wi h oluene. The c ude
p oduc was pu ified by column ch oma og aphy (2:3 E OAc–
hexane).
4.6. Gene al p ocedu e II o he syn hesis o subs i u ed
2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-glucopy anosyl)-
320
benzimidazoles om O-pe benzoyla ed e hyl
(C-b-
D
-glycopy anosyl) o mimida e (6)
E hyl C-(2,3,4,6- e a-O-benzoyl-b-
D
-glycopy anosyl) o mimi-
da e (6, 0.1 g, 0.15 mmol) and a 1,2-diaminobenzene (a–i,
0.30 mmol, 2 equi ) we e dissol ed in anhyd ous CH
2
Cl
2
(3 mL).
The mix u e was efluxed and moni o ed by TLC (2:3 E OAc–hex-
ane). A e comple ion o he eac ion he sol en was e apo a ed,
and he esidue was pu ified by column ch oma og aphy.
4.7. Gene al p ocedu e III o he Zemplén-deacyla ion
A benzoyla ed compound was dissol ed in d y MeOH (5 mL/
330
100 mg, a ew d ops o CHCl
3
we e added in case o incomple e dis-
solu ion) and a ca aly ic amoun o a NaOMe solu ion (1M in
MeOH) was added. The mix u e was kep a and moni o ed by
TLC (7:3 CHCl
3
–MeOH). When he s a ing ma e ial was consumed
he mix u e was neu alised wi h a ca ion exchange esin Ambe -
lys 15 (H
+
o m), hen he esin was fil e ed o and he sol en e-
mo ed. The esidue was pu ified by column ch oma og aphy.
4.8. 2-(2
0
,3
0
,4
0
,6
0
-Te a-O-benzoyl-b-
D
-glucopy anosyl)-
benzimidazole (8a)
A: F om acid 5(0.1 g, 0.16 mmol) and o-phenylenediamine (a,
340
0.02 g, 0.16 mmol) acco ding o Gene al p ocedu e I. Yield: 0.05 g
(45%).
B: F om imida e 6(0.10 g, 0.15 mmol) and o-phenylenediamine
(a, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion
ime: 3 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane)
o yield 0.09 g (89%) o whi e solid. Mp: 122–124 °C (Li .
22
mp:
120–123 °C).
1
H and
13
C NMR da a co espond o he epo ed
spec a.
22
4.9. 5(6)-Fluo o-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
glucopy anosyl)-benzimidazole (8b) and 5(6)-fluo o-2-(3
0
,4
0
,6
0
-
350
i-O-benzoyl-2
0
-deoxy-
D
-a abino-hex-1-enopy anosyl)-
benzimidazole (9b)
F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-fluo o-
benzene (b, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II.
Reac ion ime: 5 h. Pu ified by column ch oma og aphy (1:2, hen
2:3 E OAc–hexane) o gi e 9b as he fi s han 8b as he second
ac ion.
Compound 8b: Yield: 0.051 g (46%) pale yellow solid; R
: 0.31
(2:3 E OAc–hexane); mp: 124–126 °C; [
a
]
D
=39 (c0.5, DMSO);
1
H NMR (CDCl
3
)d(ppm): 11.58 (1H, b s, benzimidazole NH),
360
7.96–6.76 (23H, m, a oma ics), 6.30–6.21, 6.02 (3 1H, 3 pseudo
, J= 9.2, 9.2 Hz in each, H-2
0
, H-3
0
, H-4
0
), 5.36 (1H, d, J= 9.2 Hz,
H-1
0
), 4.69 (1H, dd, J= 12.3, <1 Hz, H-6
0
a), 4.57 (1H, dd, J= 12.3,
5.3 Hz, H-6
0
b), 4.52 (1H, ddd, J= 9.2, 5.3, <1 Hz, H-5
0
);
13
C NMR
(DMSO-d
6
)d(ppm): 165.4, 165.1, 164.8, 164.3 (CO), 158.9
(d,
1
J
C-F
= 237 Hz, benzimidazole), 158.2 (d,
1
J
C-F
= 237 Hz, benz-
imidazole), 151.1, 150.3 (benzimidazole), 142.8 (d,
3
J
C-F
= 9 Hz,
benzimidazole), 139.0 (b s, benzimidazole), 134.2 (d,
3
J
C-F
= 9 Hz,
benzimidazole), 133.7–133.3 (a oma ics), 130.9 (b s, benzimid-
azole), 129.3–128.5 (a oma ics), 120.0 (d,
3
J
C-F
= 7 Hz, benzimid-
370
azole), 112.3 (d,
3
J
C-F
= 6 Hz, benzimidazole), 110.9 (d,
2
J
C-F
= 23 Hz,
benzimidazole), 109.7 (
2
J
C-F
= 23 Hz, benzimidazole), 104.5
(d,
2
J
C-F
= 24 Hz, benzimidazole), 98.0 (d,
2
J
C-F
= 27 Hz, benzimid-
azole), 79.1, 74.8, 74.2, 73.4, 71.5, 69.0 (C-1
0
–C-5
0
), 62.8 (C-6
0
). Anal.
Calcd o C
41
H
31
FN
2
O
9
(714.69): C, 68.90; H, 4.37; N, 3.92. Found: C,
68.84; H, 4.29; N, 4.03.
Compound 9b: Yield: 0.032 g (35%), pale yellow sy up; R
: 0.58
(2:3 E OAc–hexane); [
a
]
D
=7(c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d
(ppm): 10.41 (1H, b s, benzimidazole NH), 8.02–6.94 (18H, m, a o-
ma ics), 6.43 (1H, b s, H-2
0
), 5.95–5.92 (2H, m, H-3
0
, H-4
0
), 4.88
380
(1H, ddd, J= 4.0, <1 Hz, H-5
0
), 4.79 (2H, s, H-6
0
a, H-6
0
b);
13
CNMR
(CDCl
3
)d(ppm): 166.2, 165.6, 165.0 (CO), 159.8 (d,
1
J
C-F
= 239 Hz,
benzimidazole C-5), 146.8 (benzimidazole C-2), 145.0 (C-1
0
),
143.1, 139.8 (2 b s, benzimidazole C-3a, C-7a), 133.5–128.4
(a oma ics), 120.2 (b s, benzimidazole C-4 o C-7), 111.9
(d,
2
J
C-F
= 25 Hz, benzimidazole C-6), 104.4 (b s, benzimidazole
C-4 o C-7), 99.4 (C-2
0
), 75.2, 67.9, 67.3 (C-3
0
–C-5
0
), 61.7 (C-6
0
).
Analysis: C
34
H
25
FN
2
O
7
(592.57), ESI-MS (posi i e mode) m/z:
615.157 [M+Na]
+
, 1207.327 [2 M+Na]
+
.
4.10. 5(6)-Chlo o-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
390
glucopy anosyl)-benzimidazole (8c) and 5(6)-chlo o-2-(3
0
,4
0
,6
0
-
i-O-benzoyl-2
0
-deoxy-
D
-a abino-hex-1-enopy anosyl)-
benzimidazole (9c)
F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-chlo o-
benzene (c, 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II.
Reac ion ime: 5 h. Pu ified by column ch oma og aphy (2:3
E OAc–hexane) o gi e 9c as he fi s and 8c as he second ac ion.
Compound 8c: Yield: 0.085 g (76%), pale yellow solid; R
: 0.29
(2:3 E OAc–hexane); mp: 123–125 °C; [
a
]
D
=61 (c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d(ppm): 11.33 (1H, b s, benzimidazole NH),
400
7.94–6.95 (23H, m, a oma ics), 6.21, 6.14, 6.00 (3 1H, 3 pseudo
, J= 9.4, 9.4 Hz in each, H-2
0
, H-3
0
, H-4
0
), 5.31 (1H, d, J= 9.4 Hz,
H-1
0
), 4.72 (1H, dd, J= 12.3, <1 Hz, H-6
0
a), 4.60 (1H, dd, J= 12.3,
4.4 Hz, H-6
0
b), 4.50 (1H, ddd, J= 9.4, 4.4, <1 Hz, H-5
0
);
13
CNMR
(CDCl
3
)d(ppm): 166.5, 166.0, 165.1 (2) (CO), 149.8 (benzimidazole
C-2), 142.0, 137.8 (2 b s, benzimidazole C-3a, C-7a), 133.3–127.8
(a oma ics), 126.9, 123.3, 119.3 (b s), 112.4 (b s) (benzimidazole
C-4–C-7), 77.0, 75.5, 74.1, 71.7, 69.5 (C-1
0
–C-5
0
), 63.4 (C-6
0
). Anal.
Calcd o C
41
H
31
ClN
2
O
9
(731.15): C, 67.35; H, 4.27; N, 3.84. Found:
C, 67.47; H, 4.13; N, 3.90.
410
Compound 9c: Yield: 0.022 g (24%), pale yellow sy up; R
: 0.59
(2:3 E OAc–hexane); [
a
]
D
=5(c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d
(ppm): 11.40 (1H, b s, benzimidazole NH), 8.00–7.07 (18H, m, a o-
ma ics), 6.45 (1H, d, J= 4.0 Hz, H-2
0
), 5.98 (1H, pseudo , J= 6.6,
5.9 Hz, H-3
0
o H-4
0
), 5.94 (1H, pseudo , J= 5.9, 4.0 Hz, H-3
0
o
H-4
0
), 4.81 (1H, ddd, J= 6.6, 4.6, 4.0 Hz, H-5
0
), 4.71–4.70 (2H, m,
H-6
0
a, H-6
0
b);
13
C NMR (CDCl
3
)d(ppm): 166.1, 165.6, 164.9 (CO),
146.8 (b s, benzimidazole C-2), 145.0 (C-1
0
), 144.1, 141.8 (2 b s,
benzimidazole C-3a, C-7a), 133.6–133.2 (a oma ics), 132.0 (b s,
benzimidazole C-5), 129.8–128.3 (a oma ics), 123.9, 120.3, 119.1,
420
112.2, 111.3 (b s o each, benzimidazole C-4, C-6, C-7), 99.8
(C-2
0
), 75.2, 68.1, 67.3 (C-3
0
–C-5
0
), 61.7 (C-6
0
). Analysis:
C
34
H
25
ClN
2
O
7
(609.02), ESI-MS (posi i e mode) m/z:631.123
[M+Na]
+
, 1239.257 [2M+Na]
+
.
4.11. 5(6)-B omo-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
glucopy anosyl)-benzimidazole (8d) and 5(6)-b omo-2-(3
0
,4
0
,6
0
-
i-O-benzoyl-2
0
-deoxy-
D
-a abino-hex-1-enopy anosyl)-
benzimidazole (9d)
F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-b omo-
benzene (d, 0.06 g, 0.30 mmol) acco ding o Gene al P ocedu e II.
430
Reac ion ime: 3 h. Pu ified by column ch oma og aphy (2:3
E OAc–hexane) o gi e 9d as he fi s and 8d as he second ac ion.
Q5
É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx 5
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011
Compound 8d: Yield: 0.064 g (54%) pale yellow solid; R
: 0.31
(2:3 E OAc–hexane); mp: 126–128 °C; [
a
]
D
=62 (c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d(ppm): 8.02–6.76 (23H, m, a oma ics), 6.42,
6.29, 6.14 (3 1H, 3 pseudo , J= 9.6, 9.6 Hz in each, H-2
0
, H-3
0
,
H-4
0
), 5.39 (1H, d, J= 9.6 Hz, H-1
0
), 4.72 (1H, dd, J= 12.3, 2.2 Hz,
H-6
0
a), 4.63 (1H, dd, J= 12.3, 4.9 Hz, H-6
0
b), 4.56 (1H, ddd, J= 9.6,
4.9, 2.2 Hz, H-5
0
);
13
C NMR (CDCl
3
)d(ppm): 166.4, 166.0, 165.0
(2) (CO), 149.6 (benzimidazole C-2), 140.9, 138.1 (2 b s, benzimid-
440
azole C-3a, C-7a), 133.3–127.7 (a oma ics), 125.9, 119.0 (b s),
117.3 (b s), 115.9 (benzimidazole C-4, C-5, C-6, C-7), 77.0, 75.5,
74.1, 71.7, 69.4 (C-1
0
–C-5
0
), 63.4 (C-6
0
). Analysis: C
41
H
31
B N
2
O
9
(775.60), ESI-MS (posi i e mode) m/z:799.112 [M+Na]
+
,
1573.242 [2M+Na]
+
.
Compound 9d: Yield: 0.029 g (29%) pale yellow sy up; R
: 0.61
(2:3 E OAc–hexane); [
a
]
D
=4(c0.25, CHCl
3
);
1
H NMR (CDCl
3
)d
(ppm): 10.06 (1H, b s, imidazole NH), 8.04–7.33 (18H, m, a oma -
ics), 6.46 (1H, b s, H-2
0
), 5.97–5.93 (2H, m, H-3
0
, H-4
0
), 4.91 (1H,
ddd, J= 4.0, <1 Hz, H-5
0
), 4.81 (2H, s, H-6
0
a, H-6
0
b). Analysis:
450
C
34
H
25
B N
2
O
7
(653.48), ESI-MS (posi i e mode) m/z:677.074
[M+Na]
+
, 1329.197 [2M+Na]
+
.
4.12. 5(6)-Ni o-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
glucopy anosyl)-benzimidazole (8e) and 5(6)-ni o-2-(3
0
,4
0
,6
0
-
i-O-benzoyl-2
0
-deoxy-
D
-a abino-hex-1-enopy anosyl)-
benzimidazole (9e)
F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4-ni o-
benzene (e, 0.05 g, 0.30 mmol) acco ding o Gene al p ocedu e II.
in anhyd ous e hanol. Reac ion ime: 1 day. Pu ified by column
ch oma og aphy (1:2 E OAc–hexane) o gi e 9e as he fi s and
460
8e as he second ac ion.
Compound 8e: Yield: 0.052 g (46%) yellow solid; R
: 0.25 (2:3
E OAc–hexane); mp: 134–136 °C; [
a
]
D
=65 (c0.5, CHCl
3
);
1
H
NMR (CDCl
3
)d(ppm): 12.87 (1H, b s, benzimidazole NH), 8.45–
6.96 (23H, m, a oma ics), 6.20, 6.12, 6.01 (3 1H, 3 pseudo ,
J= 9.4, 9.4 Hz in each, H-2
0
, H-3
0
, H-4
0
), 5.33 (1H, d, J= 9.4 Hz,
H-1
0
), 4.77 (1H, dd, J= 11.8, <1 Hz, H-6
0
a), 4.63 (1H, dd, J= 11.8,
<1 Hz, H-6
0
b), 4.51 (1H, ddd, J=J= 9.4, <1 Hz, H-5
0
);
13
C NMR
(CDCl
3
)d(ppm): 166.3, 165.8, 165.1, 164.8 (CO), 152.7 (benzimid-
azole C-2), 143.3 (benzimidazole C-5), 141.6, 138.0 (2 b s, benz-
470
imidazole C-3a, C-7a), 133.5–127.6 (a oma ics), 118.9, 116.1,
111.6 (b s o each, benzimidazole C-4, C-6, C-7), 77.1, 75.4,
74.0, 71.7, 69.0 (C-1
0
–C-5
0
), 63.2 (C-6
0
). Anal. Calcd o
C
41
H
31
N
3
O
11
(741.70): C, 66.39; H, 4.21; N, 5.67. Found: C, 66.26;
H, 4.13; N, 5.79.
Compound 9e:Yield: 0.019 g (20%) yellow sy up; R
: 0.39 (2:3
E OAc–hexane); [
a
]
D
=+1 (c0.5, CHCl
3
);
1
H NMR (DMSO-d
6
)d
(ppm): 13.51 (1H, b s, benzimidazole NH), 8.13–7.48 (18H, m, a o-
ma ics), 6.40 (1H, b s, H-2
0
), 6.02 (1H, pseudo , J= 5.3, 4.0 Hz, H-3
0
o H-4
0
), 5.93 (1H, pseudo , J= 6.6, 5.9 Hz, H-3
0
o H-4
0
), 5.23 (1H,
480
ddd, J= 5.9, 5.3, 3.3 Hz, H-5
0
), 4.89 (1H, dd, J= 12.6, 5.3 Hz, H-6
0
a),
4.78 (1H, dd, J= 12.6, 3.3 Hz, H-6
0
b);
13
C NMR (DMSO-d
6
)d
(ppm): 165.3, 165.0, 164.5 (CO), 150.5 (b s), 149.7 (b s), 147.5
(b s), 145.0, 143.0, 142.3 (b s), 138.7 (b s) (benzimidazole,
C-1
0
), 133.8–128.7 (a oma ics), 119.2, 118.5, 117.8, 115.2, 112.3,
108.4 (b s o each, benzimidazole), 100.5 (C-2
0
), 74.6, 67.9, 67.0
(C-3
0
–C-5
0
), 61.5 (C-6
0
). Analysis: C
34
H
25
N
3
O
9
(619.58), ESI-MS (po-
si i e mode) m/z:642.148 [M+Na]
+
, 1261.309 [2M+Na]
+
.
4.13. 5(6)-Me hyl-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
glucopy anosyl)-benzimidazole (8 )
490
A: F om acid 5(0.1 g, 0.16 mmol) and 3,4-diamino oluene ( ,
0.02 g, 0.16 mmol) acco ding o Gene al p ocedu e I. Yield: 0.06 g
(53%).
B: F om imida e 6(0.10 g, 0.15 mmol) and 3,4-diamino oluene
( , 0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion
ime: 2 h. Pu ified by column ch oma og aphy (2:3 E OAc–hexane)
o yield 0.09 g (83%) o pale yellow sy up. R
: 0.41 (1:1 E OAc–hex-
ane); [
a
]
D
=65 (c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d(ppm): 10.73 (1H,
b s, benzimidazole NH), 7.92–6.90 (23H, m, a oma ics), 6.19, 6.11,
5.96 (3 1H, 3 pseudo , J= 9.2, 9.2 Hz in each, H-2
0
, H-3
0
, H-4
0
),
500
5.32 (1H, d, J= 9.2 Hz, H-1
0
), 4.68 (1H, dd, J= 12.2, 2.6 Hz, H-6
0
a),
4.54 (1H, dd, J= 12.2, 5.3 Hz, H-6
0
b), 4.44 (1H, ddd, J= 9.2, 5.3,
2.6 Hz, H-5
0
), 2.32 (3H, s, CH
3
);
13
C NMR (CDCl
3
)d(ppm): 166.4,
165.9, 165.2 (2) (CO), 148.1 (benzimidazole C-2), 142.4, 138.8 (2
b s, benzimidazole C-3a, C-7a), 133.4–128.0 (a oma ics, benzimid-
azole C-5), 124.4, 118.9, 111.1 (b s o each, benzimidazole C-4,
C-6, C-7), 76.8, (C-1
0
), 75.3, 74.1, 71.5, 69.5 (C-1
0
–C-5
0
), 63.4
(C-6
0
), 21.5 (CH
3
). Anal. Calcd o C
42
H
34
N
2
O
9
(710.73): C, 70.98;
H, 4.82; N, 3.94. Found: C, 70.83; H, 4.96; N, 4.09.
4.14. 4(7)-Me hyl-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
510
glucopy anosyl)-benzimidazole (8g)
F om imida e 6(0.10 g, 0.15 mmol) and 2,3-diamino oluene (g,
0.04 g, 0.30 mmol) acco ding o Gene al P ocedu e II. Reac ion
ime: 5 h. Pu ified by column ch oma og aphy (1:2 E OAc–hexane)
o yield 0.087 g (79%) o yellow solid. Mp: 117–119 °C; [
a
]
D
=43
(c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d(ppm): 10.88 (1H, b s, benzimid-
azole NH), 7.96–6.88 (23H, m, a oma ics), 6.18, 6.06, 5.92 (3 1H,
3 pseudo , J= 9.2, 9.2 Hz in each, H-2
0
, H-3
0
, H-4
0
), 5.38 (1H, d,
J= 9.2 Hz, H-1
0
), 4.69 (1H, dd, J= 11.7, <1 Hz, H-6
0
a), 4.51 (1H, dd,
J= 11.7, 5.3 Hz, H-6
0
b), 4.44 (1H, ddd, J= 9.2, 5.3, <1 Hz, H-5
0
),
520
2.33 (3H, s, CH
3
);
13
C NMR (CDCl
3
)d(ppm): 166.3, 165.8, 165.3,
165.2 (CO), 147.7 (benzimidazole C-2), 142.1 (b s, benzimidazole
C-3a, C-7a), 133.4–128.0 (a oma ics), 123.1, 122.4, 116.8, 108.8
(b s o each, benzimidazole C-4–C-7), 76.8, 75.2, 73.9, 71.4, 69.5
(C-1
0
–C-5
0
), 63.3 (C-6
0
), 16.5 (CH
3
). Anal. Calcd o C
42
H
34
N
2
O
9
(710.73): C, 70.98; H, 4.82; N, 3.94. Found: C, 70.87; H, 4.90; N,
3.85.
4.15. 5,6-Dime hyl-2-(2
0
,3
0
,4
0
,6
0
- e a-O-benzoyl-b-
D
-
glucopy anosyl)-benzimidazole (8h)
A: F om acid 5(0.1 g, 0.16 mmol) and 1,2-diamino-4,5-dime h-
530
ylbenzene (h, 0.02 g, 0.16 mmol) acco ding o Gene al p ocedu e I.
Yield: 0.05 g (43%).
B: F om imida e 6(0.10 g, 0.15 mmol) and 1,2-diamino-4,5-
dime hylbenzene (h, 0.04 g, 0.30 mmol) acco ding o Gene al p o-
cedu e II. Reac ion ime: 3 h. Pu ified by column ch oma og aphy
(2:3 E OAc–hexane) o yield 0.09 g (81%) o pale yellow solid.
Mp: 207–209 °C; [
a
]
D
=64 (c0.5, CHCl
3
);
1
H NMR (CDCl
3
)d
(ppm): 11.00 (1H, b s, benzimidazole NH), 7.98–6.92 (22H, m, a o-
ma ics), 6.27, 6.21, 6.05 (3 1H, 3 pseudo , J= 9.2, 9.2 Hz in each,
H-2
0
, H-3
0
, H-4
0
), 5.34 (1H, d, J= 9.2 Hz, H-1
0
), 4.67 (1H, dd, J= 11.9,
540
2.6 Hz, H-6
0
a), 4.54 (1H, dd, J= 11.9, 5.3 Hz, H-6
0
b), 4.48 (1H, ddd,
J= 9.2, 5.3, 2.6 Hz, H-5
0
), 2.17 (6H, s, CH
3
);
13
C NMR (CDCl
3
)d
(ppm): 166.3, 165.9, 165.1, 165.0 (CO), 147.5 (benzimidazole
C-2), 141.1 (b s, benzimidazole C-3a, C-7a), 133.2–127.9 (a oma -
ics, benzimidazole C-5, C-6), 119.5, 111.6 (2 b s, benzimidazole C-
4, C-7), 76.8, 75.6, 74.3, 71.7, 69.5 (C-1
0
–C-5
0
), 63.3 (C-6
0
), 20.2
(2 CH
3
). Anal: Calcd o C
43
H
36
N
2
O
9
(724.75): C, 71.26; H, 5.01;
N, 3.87. Found: C, 71.15; H, 4.92; N, 3.99.
4.16. 2-(2
0
,3
0
,4
0
,6
0
-Te a-O-benzoyl-b-
D
-glucopy anosyl)-
naph ho[2,3-d]imidazole (8i)
550
F om imida e 6(0.10 g, 0.15 mmol) and 2,3-diamino-naph ha-
lene (i, 0.05 g, 0.30 mmol) acco ding o Gene al p ocedu e II.
6É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011
Reac ion ime: 4 h. Pu ified by column ch oma og aphy (4:5
E OAc–hexane) o yield 0.091 g (79%) o pale b own solid. Mp:
184–186 °C; [
a
]
D
=113 (c0.25, CHCl
3
);
1
H NMR (CDCl
3
)d
(ppm): 10.64 (1H, b s, naph hoimidazole NH), 7.81–6.92 (26H,
m, a oma ics), 6.14, 6.05, 5.90 (3 1H, 3 pseudo , J= 9.6, 9.6 Hz
in each, H-2
0
, H-3
0
, H-4
0
), 5.29 (1H, d, J= 9.6 Hz, H-1
0
), 4.64 (1H,
dd, J= 12.3, <1 Hz, H-6
0
a), 4.49 (1H, dd, J= 12.3, 5.3 Hz, H-6
0
b),
4.39 (1H, ddd, J= 9.6, 5.3, <1 Hz, H-5
0
);
13
C NMR (CDCl
3
)d(ppm):
560
166.2, 165.9, 165.1 (2) (CO), 152.9 (naph hoimidazole C-2), 142.7
(b s, naph hoimidazole C-3a, C-9a), 133.2–127.8, 123.6 (a oma -
ics), 116.2, 107.8 (2 b s, naph hoimidazole C-4, C-9), 76.9, 75.6,
74.2, 71.7, 69.5 (C-1
0
–C-5
0
), 63.4 (C-6
0
). Analysis: C
45
H
34
N
2
O
9
(746.76), ESI-MS (posi i e mode) m/z:769.219 [M+Na]
+
,
1516.457 [2M+Na]
+
.
4.17. 5(6)-Fluo o-2-(b-
D
-glucopy anosyl)-benzimidazole (10b)
F om 8b (0.22 g, 0.30 mmol) acco ding o Gene al P ocedu e III.
Reac ion ime: 3.5 h. Pu ified by column ch oma og aphy (4:1
CHCl
3
–MeOH) o yield 0.048 g (54%) o pale yellow sy up. R
:
570
0.19 (10:3 CHCl
3
–MeOH); [
a
]
D
=+9 (c0.2, DMSO);
1
H NMR
(CD
3
OD) d(ppm): 7.48 (1H, s, benzimidazole H-4), 7.21 (1H, d,
J= 8.7 Hz, benzimidazole H-7), 6.97 (1H, , J= 8.7 Hz, benzimid-
azole H-6), 4.46 (1H, d, J= 9.4 Hz, H-1
0
), 3.88 (1H, dd, J= 11.8,
<1 Hz, H-6
0
a), 3.73 (1H, dd, J= 11.8, <1 Hz, H-6
0
b), 3.61 (1H, pseudo
, J= 9.8, 9.8 Hz, H-2
0
o H-3
0
o H-4
0
), 3.52–3.47 (3H, m, H-2
0
and/o
H-3
0
and/o H-4
0
, H-5
0
);
13
C NMR (DMSO-d
6
)d(ppm): 158.3
(d,
1
J
C-F
= 236 Hz, benzimidazole C-5,), 153.9, 143.2, 138.9, 133.7,
130.6, 119.7, 112.0, 109.7, 104.0, 98.0 (b s o each, benzimid-
azole), 81.4, 77.8, 75.9, 72.7, 70.0 (C-1
0
–C-5
0
), 61.3 (C-6
0
). Anal.
580
Calcd o C
13
H
15
FN
2
O
5
(298.27): C, 52.35; H, 5.07; N, 9.39. Found:
C, 52.26; H, 5.21; N, 9.33.
4.18. 5(6)-Chlo o-2-(b-
D
-glucopy anosyl)-benzimidazole (10c)
F om 8c (0.24 g, 0.33 mmol) acco ding o gene al p ocedu e III.
Reac ion ime: 5 h. Pu ified by column ch oma og aphy (4:1
CHCl
3
–MeOH) o yield 0.092 g (89%) o colou less sy up. R
: 0.23
(4:1 CHCl
3
–MeOH); [
a
]
D
= +23 (c0.5, DMSO);
1
H NMR (CD
3
OD) d
(ppm): 7.50 (1H, d, J= 1.4 Hz, benzimidazole H-4), 7.45 (1H, d,
J= 8.9 Hz, benzimidazole H-7), 7.15 (1H, dd, J= 8.9, 1.4 Hz, benz-
imidazole H-6), 4.47 (1H, d, J= 8.9 Hz, H-1
0
), 3.88 (1H, dd, J= 11.6,
590
<1 Hz, H-6
0
a), 3.73 (1H, dd, J= 11.6, 4.8 Hz, H-6
0
b), 3.61 (1H, pseudo
, J= 8.9, 8.9 Hz, H-2
0
o H-3
0
o H-4
0
), 3.55–3.45 (3H, m, H-2
0
and/o
H-3
0
and/o H-4
0
, H-5
0
);
13
C NMR (CD
3
OD) d(ppm): 155.2 (benz-
imidazole C-2), 140.3, 138.1 (2 b s, benzimidazole C-3a, C-7a),
129.2 (benzimidazole C-5), 124.1, 117.1 (b s), 115.9(b s) (benz-
imidazole C-4, C-6, C-7), 82.2, 79.2, 77.3, 74.8, 71.2 (C-1
0
–C-5
0
),
62.8 (C-6
0
). Anal. Calcd o C
13
H
15
ClN
2
O
5
(314.72): C, 49.61; H,
4.80; N, 8.90. Found: C, 49.72; H, 4.87; N, 8.75.
4.19. 5(6)-B omo-2-(b-
D
-glucopy anosyl)-benzimidazole (10d)
F om 8d (0.27 g, 0.35 mmol) acco ding o Gene al p ocedu e III.
600
Reac ion ime: 6 h. Pu ified by column ch oma og aphy (8:1
CHCl
3
–MeOH) o yield 0.073 g (58%) o colou less sy up. R
: 0.23
(10:3 CHCl
3
–MeOH); [
a
]
D
= +21 (c0.43, DMSO);
1
H NMR (CD
3
OD)
d(ppm): 7.65 (1H, s, benzimidazole H-4), 7.41, 7.29 (2 1H, 2d,
J= 8.4 Hz, benzimidazole H-6, H-7), 4.47 (1H, d, J= 9.4 Hz, H-1
0
),
3.88 (1H, dd, J= 11.8, <1 Hz, H-6
0
a), 3.74 (1H, dd, J= 11.8, <1 Hz,
H-6
0
b), 3.61 (1H, pseudo , J= 9.4, 8.4 Hz, H-2
0
o H-3
0
o H-4
0
),
3.53–3.46 (3H, m, H-2
0
and/o H-3
0
and/o H-4
0
, H-5
0
);
13
C NMR
(DMSO-d
6
)d(ppm): 153.8 (benzimidazole C-2), 141.5, 134.8 (2
b s, C-3a, C-7a), 124.5, 121.0, 113.9, 113.8 (b s o each, benzimid-
610
azole C-4–C-7), 81.5, 77.8, 75.9, 72.8, 70.0 (C-1
0
–C-5
0
), 61.3 (C-6
0
).
Anal. Calcd o C
13
H
15
B N
2
O
5
(359.17): C, 43.47; H, 4.21; N, 7.80.
Found: C, 43.43; H, 4.34; N, 7.93.
4.20. 2-(b-
D
-Glucopy anosyl)-5(6)-ni o-benzimidazole (10e)
F om 8e (0.30 g, 0.40 mmol) acco ding o Gene al p ocedu e III.
Reac ion ime: 1 day. Pu ified by column ch oma og aphy (85:15
CHCl
3
–MeOH) o yield 0.07 g (53%) o yellow sy up. R
: 0.27 (4:1
CHCl
3
–MeOH);
1
H NMR (DMSO-d
6
)d(ppm): 13.03 (1H, b s, benz-
imidazole NH), 8.44 (1H, d, J= 1.3 Hz, benzimidazole H-4), 8.11
(1H, dd, J= 9.2, 1.3 Hz, benzimidazole H-6), 7.70 (1H, d, J= 9.2 Hz,
620
benzimidazole H-7), 5.14 (3H, b s, 3 OH), 4.60 (1H, b s, OH),
4.42 (1H, d, J= 9.2 Hz, H-1
0
), 3.74 (1H, dd, J= 11.9, <1 Hz, H-6
0
a),
3.63 (1H, pseudo , J= 9.2, 9.2 Hz, H-2
0
o H-3
0
o H-4
0
), 3.49 (1H,
dd, J= 11.9, 5.3 Hz, H-6
0
b), 3.39–3.34 (2H, m, H-2
0
o H-3
0
o H-4
0
,
H-5
0
), 3.24 (1H, pseudo , J= 9.2, 9.2 Hz, H-2
0
o H-3
0
o H-4
0
);
13
C
NMR (DMSO-d
6
)d(ppm): 157.3 (benzimidazole C-2), 142.5 (benz-
imidazole C-5), 141.4, 138.8 (2 b s, benzimidazole C-3a, C-7a),
117.7, 114.3 (b s), 112.6 (b s) (benzimidazole C-4, C-6, C-7),
81.6, 77.7, 76.0, 72.8, 70.0 (C-1
0
–C-5
0
), 61.2 (C-6
0
). Anal. Calcd o
C
13
H
15
N
3
O
7
(325.27): C, 48.00; H, 4.65; N, 12.92. Found: C, 47.87;
630
H, 4.56; N, 12.96.
4.21. 2-(b-
D
-Glucopy anosyl)-5(6)-me hyl-benzimidazole (10 )
F om 8e (0.27 g, 0.38 mmol) acco ding o Gene al p ocedu e III.
Reac ion ime: 3 h. Pu ified by column ch oma og aphy (85:15
CHCl
3
–MeOH) o yield 0.10 g (87%) o colou less sy up. R
: 0.42
(7:3 CHCl
3
–MeOH); [
a
]
D
= +18 (c0.5, MeOH);
1
H NMR (CD
3
OD) d
(ppm): 7.41 (1H, d, J= 7.9 Hz, benzimidazole H-6 o H-7), 7.32
(1H, s, benzimidazole H-4), 7.03 (1H, d, J= 7.9 Hz, benzimidazole
H-6 o H-7), 4.49 (1H, d, J= 9.2 Hz, H-1
0
), 3.91 (1H, dd, J= 11.9,
<1 Hz, H-6
0
a), 3.76 (1H, dd, J= 11.9, 4.0 Hz, H-6
0
b), 3.66 (1H, pseudo
640
, J= 9.2, 9.2 Hz, H-2
0
o H-3
0
o H-4
0
), 3.61–3.49 (3H, m, H-2
0
and/o
H-3
0
and/o H-4
0
, H-5
0
), 2.42 (3H, s, CH
3
);
13
C NMR (CD
3
OD) d
(ppm): 153.4 (benzimidazole C-2), 139.0, 137.6 (benzimidazole
C-3a, C-7a), 133.6 (benzimidazole C-5), 125.2, 115.9, 115.4 (benz-
imidazole C-4, C-6, C-7), 82.1, 79.3, 77.3, 74.8, 71.2 (C-1
0
–C-5
0
),
62.7 (C-6
0
), 21.7 (CH
3
). Anal. Calcd o C
14
H
18
N
2
O
5
(294.30): C,
57.13; H, 6.16; N, 9.52. Found: C, 57.29; H, 6.07; N, 9.39.
4.22. 2-(b-
D
-Glucopy anosyl)-4(7)-me hyl-benzimidazole (10g)
F om 8g (0.25 g, 0.35 mmol) acco ding o Gene al P ocedu e III.
Reac ion ime: 4 h. Pu ified by column ch oma og aphy (4:1
650
CHCl
3
–MeOH) o gi e 0.075 g (73%) o colou less sy up. R
: 0.20
(4:1 CHCl
3
–MeOH); [
a
]
D
= +13 (c0.5, DMSO);
1
H NMR (DMSO-d
6
)
d(ppm): 12.46 (1H, b s, benzimidazole NH), 7.32, 7.04, 6.95
(3 1H, benzimidazole), 5.14 (3H, b s, 3 OH), 4.64 (1H, b s,
OH), 4.36 (1H, d, J= 10.2 Hz, H-1
0
), 3.73–3.24 (6H, m, H-2
0
, H-3
0
,
H-4
0
, H-5
0
, H-6
0
a, H-6
0
b), 2.50 (3H, s, CH
3
);
13
C NMR (DMSO-d
6
)d
(ppm): 151.9 (benzimidazole C-2), 141.8, 133.5, 128.4, 121.6,
115.6, 109.0 (b s o each, benzimidazole C-3a, C-7a, C-4–C-7),
81.3, 77.9, 75.9, 72.7, 70.0 (C-1
0
–C-5
0
), 61.2 (C-6
0
). Anal. Calcd o
C
14
H
18
N
2
O
5
(294.30): C, 57.13; H, 6.16; N, 9.52. Found: C, 57.18;
660
H, 6.05; N, 9.64.
4.23. 5,6-Dime hyl-2-(b-
D
-glucopy anosyl)-benzimidazole (10h)
F om 8h (0.27 g, 0.37 mmol) acco ding o Gene al p ocedu e III.
Reac ion ime: 3 h. Pu ified by column ch oma og aphy (85:15
CHCl
3
–MeOH) o yield 0.09 g (78%) o colou less sy up. R
: 0.42
(7:3 CHCl
3
–MeOH); [
a
]
D
= +24 (c0.5, MeOH);
1
H NMR (CD
3
OD) d
(ppm): 7.28 (2H, s, benzimidazole H-4, H-7), 4.47 (1H, d,
J= 9.2 Hz, H-1
0
), 3.91 (1H, dd, J= 11.9, <1 Hz, H-6
0
a), 3.76 (1H, dd,
É a Boko e al. /Ca bohyd a e Resea ch xxx (2013) xxx–xxx 7
CAR 6384 No. o Pages 9, Model 5G
5 Feb ua y 2013
Please ci e his a icle in p ess as: Boko ,É.; e al. Ca bohyd . Res. (2013), h p://dx.doi.o g/10.1016/j.ca es.2013.01.011